August 15, 2024 longcha9

Determination of the content of four cyclohexene ether terpenes in Morinda officinalis from different origins using the one test multiple evaluation method
Morinda officinalis, derived from the dried roots of the Rubiaceae plant Morinda officinalis How. This traditional Chinese medicine was first recorded in the “Shennong Bencao Jing” and is classified as a top-grade herb. It has been recorded throughout history and is a famous health Chinese medicine. It is also one of the “Four Great Southern Medicines” in China, with Guangdong as the local production area. It has effects such as anti osteoporosis, tonifying kidney yang, antioxidation, anti-tumor, anti-inflammatory, anti fatigue, and enhancing the body’s immunity. The main components of Morinda officinalis include iridoids, anthraquinones, sugars, and organic acids. The iridoid compounds represented by crystal orchid glycosides have anti-inflammatory, analgesic, antioxidant, anti-tumor, anti-aging, and anti obesity effects. Caryophyllin also has bone protective effects and reduces acute lung injury.

In recent years, as people’s awareness has shifted from medical care to preventive healthcare, the demand for Morinda officinalis has continued to increase, gradually leading to the phenomenon of substituting inferior for good. The 2015 edition of the Chinese Pharmacopoeia uses one component, Nissin, as a quantitative indicator for the content determination of Morinda officinalis, which has certain limitations. If multiple indicator components can be included in the quantitative indicator at the same time, it will provide a more comprehensive evaluation of the quality of Morinda officinalis.

The one test multiple evaluation method is a method that utilizes the intrinsic functions and proportional relationships between the active ingredients of traditional Chinese medicine to achieve synchronous determination of multiple ingredients (difficult to obtain or expensive to obtain reference standards) by measuring one ingredient (reference standards are easy to obtain). This method is a key promotion method in the 2015 edition of the Chinese Pharmacopoeia and has become the preferred method for determining the content of multiple indicators in traditional Chinese medicine. However, there are currently no reports at home and abroad on the use of the single test multiple evaluation method to determine its content.

This experiment established a one test multiple evaluation method, which for the first time achieved synchronous determination of four cyclohexene ether terpenoid components in Morinda officinalis, including crystal orchid glycoside, deacetylated coumarinic acid, coumarinic acid, and coumarin. Quality control was also carried out on Morinda officinalis from different origins, providing reference for the revision of its quality standards.

 

This experiment investigated the extraction solvents (70%, 80%, 90% methanol), different extraction methods (ultrasonic extraction, heating reflux extraction), and different extraction times (20, 30, 40 min), taking into account various factors such as the ease of operation of the extraction method, operating costs, and the extraction rate of the tested components. Finally, the preparation method of the Morinda officinalis test solution was determined to use 80% methanol as the extraction solvent, ultrasonic extraction twice, and each extraction time of 30 min.

In this experiment, methanol phosphoric acid water and acetonitrile phosphoric acid water were used as mobile phases. It was found that when acetonitrile phosphoric acid water was used as the mobile phase, the peak shape and separation degree of each tested component were good, and the theoretical number of trays was also high. Finally, the mobile phase was determined as acetonitrile (A)~0.2% phosphoric acid solution+0.01 mol/L dipotassium dihydrogen phosphate buffer salt (B), gradient elution (0-12 min, 1% A; 12-30 min, 1% A → 17% A; 30-40 min, 17% A).

Due to the low price, easy availability, and stable properties of the reference substance for deacetylated coumarin acid, this experiment will use deacetylated coumarin acid as an internal reference for multi evaluation analysis.

The QAMS method established in this experiment achieved for the first time the simultaneous determination of four iridoid components in Morinda officinalis, including crystal orchid glycoside, deacetylated coumarinic acid, coumarinic acid, and coumarin. Through methodological investigation, relative correction factor durability experiments, and comparison of QAMS method calculated values with ESM method measured values RSD values, it is shown that the method established in this paper is easy to operate, accurate in results, and has good repeatability. Solved the shortcomings of the traditional multi index component evaluation model, such as large use of reference materials, high detection costs, and difficulty in obtaining some reference materials. This can provide a reference for the comprehensive evaluation of the quality of Morinda officinalis.

There are differences in the content of various components in Morinda officinalis from different regions, with a more significant difference in the content of crystal orchid glycosides. The content of crystal orchid glycosides in the 9th and 10th batches is lower, which may be related to the fact that Morinda officinalis comes from different regions.

In summary, the QAMS method established in this experiment is accurate, reliable, and has good reproducibility, which can provide a new method for quality control of Morinda officinalis

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